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Claudins constitute TJ strands in conjunction with the cytoplasmic scaffolds ZO-1 and ZO-2 and play pivotal roles in epithelial barrier formation. However, how claudins and other TJ membrane proteins cooperate to organize TJs remains unclear. Here, we systematically knocked out TJ components by genome editing and show that while ZO-1\/ZO-2\u2013deficient cells lacked TJ structures and epithelial barriers, claudin-deficient cells lacked TJ strands and an electrolyte permeability barrier but formed membrane appositions and a macromolecule permeability barrier. Moreover, epithelial polarity was disorganized in ZO-1\/ZO-2\u2013deficient cells, but not in claudin-deficient cells. Simultaneous deletion of claudins and a TJ membrane protein JAM-A resulted in a loss of membrane appositions and a macromolecule permeability barrier and in sporadic epithelial polarity defects. These results demonstrate that claudins and JAM-A coordinately regulate TJ formation and epithelial polarity.<\/jats:p>","DOI":"10.1083\/jcb.201812157","type":"journal-article","created":{"date-parts":[[2019,8,29]],"date-time":"2019-08-29T13:29:02Z","timestamp":1567085342000},"page":"3372-3396","update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":256,"title":["Claudins and JAM-A coordinately regulate tight junction formation and epithelial polarity"],"prefix":"10.1083","volume":"218","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2557-6722","authenticated-orcid":true,"given":"Tetsuhisa","family":"Otani","sequence":"first","affiliation":[{"name":"Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Aichi, Japan 1"},{"name":"Department of Physiological Sciences, School of Life Science, SOKENDAI (The Graduate University for Advanced Studies), Okazaki, Aichi, Japan 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8074-9588","authenticated-orcid":true,"given":"Thanh Phuong","family":"Nguyen","sequence":"additional","affiliation":[{"name":"Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Aichi, Japan 1"},{"name":"Department of Physiological Sciences, School of Life Science, SOKENDAI (The Graduate University for Advanced Studies), Okazaki, Aichi, Japan 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3726-7775","authenticated-orcid":true,"given":"Shinsaku","family":"Tokuda","sequence":"additional","affiliation":[{"name":"Division of Nephrology and Hypertension, Department of Internal Medicine, University of Kansas Medical Center, Kansas City, KS 3"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5502-6967","authenticated-orcid":true,"given":"Kei","family":"Sugihara","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Cell Biology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan 4"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0675-7859","authenticated-orcid":false,"given":"Taichi","family":"Sugawara","sequence":"additional","affiliation":[{"name":"Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Aichi, Japan 1"},{"name":"Department of Physiological Sciences, School of Life Science, SOKENDAI (The Graduate University for Advanced Studies), Okazaki, Aichi, Japan 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4304-6204","authenticated-orcid":true,"given":"Kyoko","family":"Furuse","sequence":"additional","affiliation":[{"name":"Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Aichi, Japan 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9465-275X","authenticated-orcid":true,"given":"Takashi","family":"Miura","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Cell Biology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan 4"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0417-7888","authenticated-orcid":true,"given":"Klaus","family":"Ebnet","sequence":"additional","affiliation":[{"name":"Institute-Associated Research Group \u201cCell Adhesion and Cell Polarity,\u201d Institute of Medical Biochemistry, Zentrum f\u00fcr Molekularbiologie der Entz\u00fcndung, University of M\u00fcnster, M\u00fcnster, Germany 5"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2847-8156","authenticated-orcid":true,"given":"Mikio","family":"Furuse","sequence":"additional","affiliation":[{"name":"Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Aichi, Japan 1"},{"name":"Department of Physiological Sciences, School of Life Science, SOKENDAI (The Graduate University for Advanced Studies), Okazaki, Aichi, Japan 2"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2019,8,29]]},"reference":[{"key":"2023072315145456100_bib1","doi-asserted-by":"publisher","first-page":"14867","DOI":"10.1038\/ncomms14867","article-title":"The Par3 polarity protein is an exocyst receptor essential for mammary cell survival","volume":"8","author":"Ahmed","year":"2017","journal-title":"Nat. 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